Inadequate Differentiation of Theileria orientalis Genotypes buffeli and
            <i>ikeda</i>
            in a Multiplexed Tandem PCR (MT-PCR) Assay Using the
            <i>p23</i>
            Gene as a Marker

Gebrekidan, Hagos; Gasser, Robin B.; Jabbar, Abdul

doi:10.1128/jcm.02017-16

Cited by 3 publications

(1 citation statement)

References 16 publications

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“…The majority of these assays have been designed to specifically detect the pathogenic Ikeda genotype [93,146,163,164,166] and in some cases several genotype specific assays have been multiplexed [146,163,166]. Genotype discrimination has been most successfully achieved using assays targeting the MPSP gene [146,163] while some other molecular markers have been shown to be insufficiently discriminatory [167].…”

Section: Diagnosismentioning

confidence: 99%

Oriental Theileriosis

Yam¹,

Bogema²,

Jenkins³

2019

Ticks and Tick-Borne Pathogens

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Theileria orientalis, the causative agent of oriental theileriosis, is an apicomplexan haemoparasite and is one of several tick-borne Theileria spp. infecting cattle. Unlike the highly pathogenic transforming Theileria species (T. annulata and T. parva) which induce uncontrolled lymphocytic proliferation, T. orientalis is a non-transforming strain exerting its major pathogenic effects via erythrocyte destruction. Clinical symptoms associated with oriental theileriosis are largely consequences of the underlying anaemia. Because of its non-transforming nature, T. orientalis was previously considered a benign parasite, however, in the recent years, clinical outbreaks of T. orientalis have been increasingly observed throughout Asia and Australasia. Recent rapid spread of clinical theileriosis has been linked to a pathogenic genotype of the parasite, genotype Ikeda (Type 2). The geographic distribution of clinical outbreaks correlates to the range of the major vector tick, Haemaphysalis longicornis, although other vectors and modes of transmission are possible. This review includes discussion of T. orientalis epidemiology, transmission, pathogenesis, treatment and control and provides an update on the taxonomy of this organism which is still under debate.

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Section: Diagnosismentioning

confidence: 99%

Oriental Theileriosis

Yam¹,

Bogema²,

Jenkins³

2019

Ticks and Tick-Borne Pathogens

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An appraisal of oriental theileriosis and the Theileria orientalis complex, with an emphasis on diagnosis and genetic characterisation

et al. 2019

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Oriental theileriosis, a tick-borne disease of bovids caused by members of the Theileria orientalis complex, has a worldwide distribution. Globally, at least 11 distinct genotypes of T. orientalis complex, including type 1 (chitose), type 2 (ikeda), type 3 (buffeli), types 4 to 8, and N1-N3, have been described based on the sequence of the major piroplasm surface protein (MPSP) gene. Of these 11 genotypes, mainly ikeda and chitose are known to be pathogenic and cause considerable morbidity (including high fever, anaemia, jaundice and abortion), production losses and/or mortality in cattle. Mixed infections with two or more genotypes of T. orientalis is common, but do not always lead to a clinical disease, posing challenges in the diagnosis of asymptomatic or subclinical forms of oriental theileriosis. The diagnosis of oriental theileriosis is usually based on clinical signs, the detection of piroplasms of T. orientalis in blood smears, and/or the use of serological or molecular techniques. This paper reviews current methods used for the diagnosis of T. orientalis infections and the genetic characterisation of members of the T. orientalis complex, and proposes that advanced genomic tools should be established for investigations of these and related haemoparasites.

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Multiplexed Tandem PCR (MT-PCR) Assay Using the Major Piroplasm Surface Protein Gene for the Diagnosis of Theileria orientalis Infection in Cattle

et al. 2018

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O riental theileriosis is an economically important tick-borne disease of bovines, particularly in the Asia-Pacific region, and is caused by one or more genotypes of the Theileria orientalis complex (1, 2). Based on sequence analysis of the major piroplasm surface protein (MPSP) gene, at least 11 genotypes have been defined for T. orientalis (1); five of them (i.e., Buffeli, Chitose, Ikeda, and types 4 and 5) have been reported from Australia (3, 4). Previously, high infection intensities of the pathogenic genotypes Ikeda and Chitose have been linked to the clinical cases of oriental theileriosis in Australia and New Zealand (5-7); hence, it is important to estimate the burden of different genotypes of T. orientalis. To detect, differentiate, or quantitate these genotypes, three molecular diagnostic techniques, including conventional PCR (4), quantitative PCR using TaqMan probes (5, 6), and multiplexed tandem PCR (MT-PCR) (7), have been utilized. Although a previously developed multiplexed tandem PCR (MT-PCR) for the detection, differentiation, and quantitation of four genotypes (Buffeli, Chitose, Ikeda, and type 5) of T. orientalis was sensitive, cost-effective, and time-effective (7), the direct comparison of the intensities of infection for different genotypes was challenging using the assay due to the use of multiple markers (i.e., the MPSP gene for Chitose and type 5, the p23 gene for Buffeli, and ITS-1 for Ikeda) with variable copy numbers per genome of T. orientalis (8). Furthermore, cross-reactivity was detected between genotypes Buffeli and Ikeda (using the p23 gene) when samples originating from other geographical regions such as Ethiopia and Pakistan were tested by MT-PCR (8). The objective of this study was to modify the existing MT-PCR assay to detect, differentiate, and quantitate the four common genotypes of T. orientalis (Buffeli, Chitose, Ikeda, and type 5) using the MPSP marker. A total of 175 DNA samples (known test positives [n ϭ 50] and known test negatives [n ϭ 125] by conventional PCR and DNA sequencing) extracted from cattle blood samples were available from previous studies (9, 10). The 50 test-positive-control samples were positive for at least one of the four genotypes of T. orientalis (Buffeli, Chitose, Ikeda, and type 5) (9, 10). The assay was conducted in the 24-well variant of the Easy-Plex platform, including the 96-well Easy-Plex Analyzer and PC with Easy-Plex software (catalog no. 9350; AusDiagnostics Pty. Ltd., Australia). The primary amplification ("target enrichment") was conducted using genotype-specific primer pairs designed to the sequences of the MPSP gene for genotypes Buffeli, Chitose, Ikeda, and type 5 and the first internal transcribed spacer (ITS-1) for all Theileria spp. infecting bovines (Step 1 tubes for Theileria [8 well], catalog no. 78171S; AusDiagnostics). The secondary amplification for

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Inadequate Differentiation of Theileria orientalis Genotypes buffeli and ikeda in a Multiplexed Tandem PCR (MT-PCR) Assay Using the p23 Gene as a Marker

Cited by 3 publications

References 16 publications

Oriental Theileriosis

Oriental Theileriosis

An appraisal of oriental theileriosis and the Theileria orientalis complex, with an emphasis on diagnosis and genetic characterisation

Multiplexed Tandem PCR (MT-PCR) Assay Using the Major Piroplasm Surface Protein Gene for the Diagnosis of Theileria orientalis Infection in Cattle

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